Comparison of pork belly characteristics and weights of primal cuts between gilt and barrow of Landrace × Yorkshire × Duroc pigs measured by AutoFomIII.

Currently, pigs breed in Korea are LYD (Landrace × Yorkshire × Duroc) crossbred pigs. Pigs used as fresh meat are gilts and barrows. However, the current supply of pork is not satisfying Korean consumers. In addition, the comparison of carcasses between gilts and barrows only studies carcass weight, backfat thickness, or meat quality, and there are very few studies comparing carcass characteristics. The purpose of this study was to compare characteristics of 7 primal cuts of gilts and barrows as measured by AutoFom III. A total of 350,179 pigs were used, including 176,461 gilts and 173,718 barrows. Characteristics of seven primal cuts were measured using AutoFom III. In the case of carcass weight, there was no significant difference in grade 1+. For all other survey items except for grade 2, gilts showed significantly (p < 0.05) higher values. For all grades except for pork belly, amounts of the remaining six primal cuts were higher in gilts (all p < 0.05,). In addition, the ratio of intermuscular fat in the pork belly of barrows showed a higher value than that in the pork belly of gilts (p < 0.05). The amount of pork belly, which is the most popular among consumers in Korea, not only produced more production than gilts in barrows, but also showed a higher value than gilts in barrows for the ratio of intermuscular fat affecting taste. In summary, gilts produced higher yields than barrows in all parts except pork belly. For the production of only pork belly, barrows showed higher production than gilts.

Correlation between the Korean pork grade system and the amount of pork primal cut estimated with AutoFom III.

It is impossible to know the amount of pork primal cut by pig carcass grade which is determined only by carcass weight and backfat thickness in the Korean Pig Carcass System. The aim of this study was to investigate the correlation between the pig carcass grade and the amount of pork primal cut estimated with AutoFom III. A total of 419,321 Landrace, Yorkshire, and Duroc (LYD) pigs were graded with the Korean Pig Carcass Grade System. Amounts of belly, neck, loin, tenderloin, spare ribs, shoulder, and ham were estimated with AutoFom III. Regression equations for seven primal cuts according to each grade were derived. There were significant differences among the three carcass grades due to heteroscedasticity variance (p < 0.0001). Three regression equations were derived from AutoFom III estimation of primal cuts according to carcass grades. The coefficient of determination of the regression equation was 0.941 for grade 1+, 0.982 for grade 1, and 0.993 for grade 2. Regression equations obtained from this study are suitable for AutoFom III software, a useful tool for the analysis of each pig carcass grade in the Korean Pig Carcass Grade System. The high reliability of predicting the amount of primal cut with AutoFom III is advantageous for the management of slaughterhouses to optimize their product sorting in Korea.

Critically coupled Fabry-Perot cavity with high signal contrast for refractive index sensing.

Perfect absorption at a resonance wavelength and extremely low absorption at the wavelength range of off-resonance in a one-port optical cavity is required for refractive index (RI) sensing with high signal contrast. Here, we propose and analyze an absorption-enhanced Fabry-Perot (MAFP) cavity based on a critical coupling condition in a near-infrared wavelength range. For a one-port cavity, a thick bottom Au is used as a mirror and an absorber. To achieve the critical coupling condition, a top dielectric metasurface is employed and tailored to balance the radiation coupling and the absorption coupling rates, and the one-port cavity is theoretically analyzed using temporal coupled-mode theory. We investigate two types of MAFP structures for gas and liquid. The gas MAFP cavity shows a sensitivity of ~ 1388 nm/RIU and a full-width at half-maximum of less than 0.7 nm. This MAFP cavity resolves the RI change of 5 × 10-4 with a reflectance signal margin of 50% and achieves a signal contrast of ~ 100%. The liquid MAFP cavity shows a sensitivity of ~ 996 nm/RIU when RI of liquid changes from 1.30 to 1.38. With tailoring the period of the metasurface maintaining its thickness, a signal contrast of ~ 100% is achieved for each specific RI range.

Tunable dual-wavelength absorption switch with graphene based on an asymmetric guided-mode resonance structure.

We propose a tunable dual-wavelength absorption (TDWA) switch based on an asymmetric guided mode resonance (AGMR) structure. A TDWA switch consists of a graphene layer and an AGMR structure sandwiched by cap and slab layers on a buffer/silicon substrate. The AGMR structure adds a smaller grating unit cell next to a larger one, exciting a second resonance close to but distinct from the first resonance. For switching, the TDWA between an absorptive or reflective mode with each on-/off-state, the chemical potential of graphene is tuned from 0.0 eV to 0.6 eV. For the absorptive mode, two absorption peaks of ≥ 96.2% are separated by 23 nm, both having an on-off ratio of ∼15.52. For the reflective mode, two reflectance peaks of ≥ 93.8% are separated by 23 nm, having on-off ratios of 15.56 dB and 18.95 dB. The maximum on-off ratios of 39.98 dB and 34.55 dB are achieved near the reflectance peaks. Both the period of the AGMR and the cap thickness alters the two peak wavelengths linearly, while the grating width of the AGMR varies nonlinearly from 17 nm to 28 nm. The buffer excites a weak Fabry-Perot resonance, which interacts with the TDWA structure, the result of which is the two absorption peaks are varied. Finally, as the incidence angle of light increases up to 5.3°, the distance of the two peak wavelengths is tuned from ∼22 nm to ∼77 nm with ≥ 96% absorption or ≥ 93% reflectance in each mode.

Dual-guiding-layer resonance structure with an embedded metasurface for quasi-critical coupling without a perfect mirror.

We propose an all-dielectric quasi-one-port resonance structure that achieves near perfect absorption without the use of a back mirror. The structure mainly consists of a high-refractive-index silicon metasurface and surrounding high-refractive-index guiding layers. The dual-guiding-layer (DGL) structure has high background reflectance and is designed to have a ratio of two decay rates into the upper and lower regions within a wider range. When an absorbing material is introduced into a DGL system, it can be designed to achieve a near critical-coupling condition by reducing the constraints in the two decay rates. By using single-layer graphene as an absorbing material, the DGL resonance structure shows an absorption of ~ 97% and a phase change of ∼ 0.95π near the wavelength of 1550 nm, confirming quasi-critical coupling. The optimized DGL structure is relatively insensitive to potential fabrication imperfections, and consequently, the expected average peak wavelength and absorption are obtained as 1549.29 nm and 96.74%, respectively. Angle-dependent absorption confirms that maximum absorption occurs under normal incidence. The DGL absorber is also designed to cover the whole C-band region, in order to meet the quasi-critical-coupling condition. All mode profiles are similarly quasi-symmetric along the metasurface due to the same DGL resonance mechanism.

Mapping the structural, electrical, and optical properties of hydrothermally grown phosphorus-doped ZnO nanorods for optoelectronic device applications.

The phosphorus-doped ZnO nanorods were prepared using hydrothermal process, whose structural modifications as a function of doping concentration were investigated using X-ray diffraction. The dopant concentration-dependent enhancement in length and diameter of the nanorods had established the phosphorus doping in ZnO nanorods. The gradual transformation in the type of conductivity as observed from the variation of carrier concentration and Hall coefficient had further confirmed the phosphorus doping. The modification of carrier concentration in the ZnO nanorods due to phosphorus doping was understood on the basis of the amphoteric nature of the phosphorus. The ZnO nanorods in the absence of phosphorus showed the photoluminescence (PL) in the range of the ultraviolet (UV) and visible regimes. The UV emission, i.e. near-band-edge emission of ZnO, was found to be red-shifted after the doping of phosphorus, which was attributed to donor-acceptor pair formation. The observed emissions in the visible regime were due to the deep level emissions that were aroused from various defects in ZnO. The Al-doped ZnO seed layer was found to be responsible for the observed near-infrared (NIR) emission. The PL emission in UV and visible regimes can cover a wide range of applications from biological to optoelectronic devices.

A methodological review on material growth and synthesis of solar-driven water splitting photoelectrochemical cells.

As a renewable and sustainable energy source and an alternative to fossil fuels, solar-driven water splitting with photoelectrochemical (PEC) cell is a promising approach to obtain hydrogen fuel with its near-zero carbon emission pathway by transforming incident sunlight, the most abundant energy source. Because of its importance and future prospects, a number of architectures with their own features have been formed by various synthesis and growth methods. Because the materials themselves are one of the most dominant components, they determine the solar-to-hydrogen efficiency of the PEC cells. Thus, several representative PEC cells were reviewed by categorizing them as per synthesis and/or growth methods such as physical vapor deposition, chemical vapor deposition, electrochemical deposition, etc. This review provides researchers with an overview and acts as a guide for research on solar-driven water splitting PEC cells.

Tailoring Heterovalent Interface Formation with Light.

Integrating different semiconductor materials into an epitaxial device structure offers additional degrees of freedom to select for optimal material properties in each layer. However, interfaces between materials with different valences (i.e. III-V, II-VI and IV semiconductors) can be difficult to form with high quality. Using ZnSe/GaAs as a model system, we explore the use of ultraviolet (UV) illumination during heterovalent interface growth by molecular beam epitaxy as a way to modify the interface properties. We find that UV illumination alters the mixture of chemical bonds at the interface, permitting the formation of Ga-Se bonds that help to passivate the underlying GaAs layer. Illumination also helps to reduce defects in the ZnSe epilayer. These results suggest that moderate UV illumination during growth may be used as a way to improve the optical properties of both the GaAs and ZnSe layers on either side of the interface.

Biallelic modification of IL2RG leads to severe combined immunodeficiency in pigs.

BACKGROUND: Pigs with SCID can be a useful model in regenerative medicine, xenotransplantation, and cancer cell transplantation studies. Utilizing genome editing technologies such as CRISPR/Cas9 system allows us to generate genetically engineered pigs at a higher efficiency. In this study, we report generation and phenotypic characterization of IL2RG knockout female pigs produced through combination of CRISPR/Cas9 system and SCNT. As expected, pigs lacking IL2RG presented SCID phenotype. METHODS: First, specific CRISPR/Cas9 systems targeting IL2RG were introduced into developing pig embryos then the embryos were transferred into surrogates. A total of six fetuses were obtained from the embryo transfer and fetal fibroblast cell lines were established. Then IL2RG knockout female cells carrying biallelic genetic modification were used as donor cells for SCNT, followed by embryo transfer. RESULTS: Three live cloned female piglets carrying biallelic mutations in IL2RG were produced. All cloned piglets completely lacked thymus and they had a significantly reduced level of mature T, B and NK cells in their blood and spleen. CONCLUSIONS: Here, we generated IL2RG knockout female pigs showing phenotypic characterization of SCID. This IL2RG knockout female pigs will be a promising model for biomedical and translational research.

Partial loss of interleukin 2 receptor gamma function in pigs provides mechanistic insights for the study of human immunodeficiency syndrome.

In this study, we described the phenotype of monoallelic interleukin 2 receptor gamma knockout (mIL2RG+/Δ69-368 KO) pigs. Approximately 80% of mIL2RG+/Δ69-368 KO pigs (8/10) were athymic, whereas 20% (2/10) presented a rudimentary thymus. The body weight of IL2RG+/Δ69-368KO pigs developed normally. Immunological analysis showed that mIL2RG+/Δ69-368 KO pigs possessed CD25+CD44- or CD25-CD44+ cells, whereas single (CD4 or CD8) or double (CD4/8) positive cells were lacking in mIL2RG+/Δ69-368 KO pigs. CD3+ cells in the thymus of mIL2RG+/Δ69-368 KO pigs contained mainly CD44+ cells and/or CD25+ cells, which included FOXP3+ cells. These observations demonstrated that T cells from mIL2RG+/Δ69-368 KO pigs were able to develop to the DN3 stage, but failed to transition toward the DN4 stage. Whole-transcriptome analysis of thymus and spleen, and subsequent pathway analysis revealed that a subset of genes differentially expressed following the loss of IL2RG might be responsible for both impaired T-cell receptor and cytokine-mediated signalling. However, comparative analysis of two mIL2RG+/Δ69-368 KO pigs revealed little variability in the down- and up-regulated gene sets. In conclusion, mIL2RG+/Δ69-368 KO pigs presented a T-B+NK- SCID phenotype, suggesting that pigs can be used as a valuable and suitable biomedical model for human SCID research.

Production of α1,3-galactosyltransferase targeted pigs using transcription activator-like effector nuclease-mediated genome editing technology.

Recent developments in genome editing technology using meganucleases demonstrate an efficient method of producing gene edited pigs. In this study, we examined the effectiveness of the transcription activator-like effector nuclease (TALEN) system in generating specific mutations on the pig genome. Specific TALEN was designed to induce a double-strand break on exon 9 of the porcine α1,3-galactosyltransferase (GGTA1) gene as it is the main cause of hyperacute rejection after xenotransplantation. Human decay-accelerating factor (hDAF) gene, which can produce a complement inhibitor to protect cells from complement attack after xenotransplantation, was also integrated into the genome simultaneously. Plasmids coding for the TALEN pair and hDAF gene were transfected into porcine cells by electroporation to disrupt the porcine GGTA1 gene and express hDAF. The transfected cells were then sorted using a biotin-labeled IB4 lectin attached to magnetic beads to obtain GGTA1 deficient cells. As a result, we established GGTA1 knockout (KO) cell lines with biallelic modification (35.0%) and GGTA1 KO cell lines expressing hDAF (13.0%). When these cells were used for somatic cell nuclear transfer, we successfully obtained live GGTA1 KO pigs expressing hDAF. Our results demonstrate that TALEN-mediated genome editing is efficient and can be successfully used to generate gene edited pigs.

Directional Terahertz Radiation from GalnP Lateral Superlattice.

We devised directionally controllable THz emission sources based on lateral composition modulation (LCM) structures. LCM structures were composed of In-rich Ga0.47In0.53P and Ga-rich Ga0.51In0.49P layers whose period was in quantum scale of ~`5 nm. The inherent type II band alignment in these structures leads to electron-hole (e-h) separation and plays a key role in generating later- ally polarized dipole ensembles, thus concomitantly emitting enhanced transmissive THz waves as compared to bulk sample. On the other hand, in lateral geometry, changes in THz fields between LCM and bulk structures turned out to negligible since the vertical electronic diffusion was allowed in both samples.

Pig oocyte activation using a Zn²âº chelator, TPEN.

Artificial oocyte activation is a critical step during SCNT. Most current activation protocols focus on inducing an increase in the intracellular free Ca(2+) concentration of the oocyte. Here, we have used a zinc chelator, TPEN, to enhance the efficiency of oocyte activation during SCNT. TPEN treatment of matured pig oocytes resulted in the reduction of available Zn(2+) in pig oocytes; however, the cytosolic Ca(2+) concentration in the oocytes was not affected by the TPEN treatment. When various concentrations (100-250 µM) and incubation durations (45 minutes-2.5 hours) of TPEN were used to activate oocytes, the efficiency of oocyte activation was not different from conventional activation methods. When oocytes that were activated by conventional activation methods were incubated with a lower concentration of TPEN (5-10 µM), a significant increase in embryos developing to the blastocyst stage was observed. In addition, when oocytes receiving a small Ca(2+) stimulus were further activated by higher concentration of TPEN (100-200 µM), a significant increase in the frequency of blastocyst formation was observed, compared to a conventional activation method. This result indicated that TPEN can be a main reagent in oocyte activation. No increase in the cytosolic Ca(2+) level was detected when oocytes were exposed to various concentrations of TPEN, indicating the ability of TPEN to induce oocyte activation is independent of an intracellular Ca(2+) increase. We were able to produce clones through SCNT by using the TPEN-assisted activation procedure, and the piglets produced through the process did not show any signs of abnormality. In this study, we have developed an efficient way to use TPEN to increase the developmental potential of cloned embryos.

Engraftment of human iPS cells and allogeneic porcine cells into pigs with inactivated RAG2 and accompanying severe combined immunodeficiency.

Pigs with severe combined immunodeficiency (SCID) may provide useful models for regenerative medicine, xenotransplantation, and tumor development and will aid in developing therapies for human SCID patients. Using a reporter-guided transcription activator-like effector nuclease (TALEN) system, we generated targeted modifications of recombination activating gene (RAG) 2 in somatic cells at high efficiency, including some that affected both alleles. Somatic-cell nuclear transfer performed with the mutated cells produced pigs with RAG2 mutations without integrated exogenous DNA. Biallelically modified pigs either lacked a thymus or had one that was underdeveloped. Their splenic white pulp lacked B and T cells. Under a conventional housing environment, the biallelic RAG2 mutants manifested a "failure to thrive" phenotype, with signs of inflammation and apoptosis in the spleen compared with age-matched wild-type animals by the time they were 4 wk of age. Pigs raised in a clean environment were healthier and, following injection of human induced pluripotent stem cells (iPSCs), quickly developed mature teratomas representing all three germ layers. The pigs also tolerated grafts of allogeneic porcine trophoblast stem cells. These SCID pigs should have a variety of uses in transplantation biology.

Cell cycle synchronization of leukemia inhibitory factor (LIF)-dependent porcine-induced pluripotent stem cells and the generation of cloned embryos.

Nuclear transfer (NT) from porcine iPSC to create cloned piglets is unusually inefficient. Here we examined whether such failure might be related to the cell cycle stage of donor nuclei. Porcine iPSC, derived here from the inner cell mass of blastocysts, have a prolonged S phase and are highly sensitive to drugs normally used for synchronization. However, a double-blocking procedure with 0.3 µM aphidicolin for 10 h followed by 20 ng/ml nocodazole for 4 h arrested 94.3% of the cells at G2/M and, after release from the block, provided 70.1% cells in the subsequent G1 phase without causing any significant loss of cell viability or pluripotent phenotype. Nuclei from different cell cycle stages were used as donors for NT to in vitro-matured metaphase II oocytes. G2/M nuclei were more efficient than either G1 and S stage nuclei in undergoing first cleavage and in producing blastocysts, but all groups had a high incidence of chromosomal/nuclear abnormalities at 2 h and 6 h compared with non-synchronized NT controls from fetal fibroblasts. Many G2 embryos extruded a pseudo-second polar body soon after NT and, at blastocyst, tended to be either polyploid or diploid. By contrast, the few G1 blastocysts that developed were usually mosaic or aneuploid. The poor developmental potential of G1 nuclei may relate to lack of a G1/S check point, as the cells become active in DNA synthesis shortly after exit from mitosis. Together, these data provide at least a partial explanation for the almost complete failure to produce cloned piglets from piPSC.

Dynamics of TET family expression in porcine preimplantation embryos is related to zygotic genome activation and required for the maintenance of NANOG.

Dynamic changes in DNA methylation are observed during embryo development. Recent studies show that the TET family is involved in these changes by converting 5-methylcytosine (5mec) to 5-hydroxymethylcytosine (5hmec). Specifically, TET3 is responsible for the conversion in the early stages, and then TET1 is a key regulator at later stages of embryo development. From previous mouse reports and our preliminary data in porcine embryos, we hypothesized that TET1 becomes the main regulator at the time of the maternal to zygotic transition (MZT). Transcript abundance of TET3 was high only at the zygote and 2-cell stage. The abundance of TET1 mRNA was high in the blastocysts and TET1 protein was present at the 4-cell stage and the blastocysts. The dynamic was similar in porcine somatic cell nuclear transfer (SCNT) embryos however; abnormally upregulated TET3 was detected at the 4-cell stage. When transcription or translation was blocked at the 2-cell stage, TET3 mRNA remained high at the 4-cell stage suggesting that degradation of TET3 is related to the MZT. Downregulation of TET3 before fertilization resulted in the reduction of 5hmec in zygotes indicating that TET3 is a key molecule for 5hmec synthesis. This misregulation of 5hmec in zygotes also affected the level of NANOG expression in the blastocysts. We show here that the porcine TET family shows dynamic expression patterns during embryogenesis, and is responsible for the appearance of 5hmec in the zygotes by TET3. This appearance of 5hmec in zygote is important for the expression of NANOG in the blastocysts.

Production of biallelic CMP-Neu5Ac hydroxylase knock-out pigs.

After the knock-out (KO) of α1,3 galactosyltransfease (Gal-T), the Hanganutziu-Deicher antigen became a major antigen of the "non-Gal antigen" that is implicated in subsequent xenograft rejection. For deletion of non-Gal antigen, we successfully produced zinc finger nuclease (ZFN)-mediated monoallelic/biallelic male and female CMP-N-acetylneuraminic acid hydroxylase (CMAH) KO miniature pigs: the efficiency of the gene targeting (41.7%) was higher when donor DNA was used with the ZFN than those of ZFN alone (9.1%). Monoallelic KO pigs had no integration of exogenous DNA into their genome, indicating that this technique would provide a new avenue to reduce the risk of antibiotics resistance when organs from genetically modified pigs are transplanted into patients. Until now, both monoallelic and biallelic CMAH KO pigs are healthy and show no sign of abnormality and off-target mutations. Therefore, these CMAH null pigs on the Gal-T KO background could serve as an important model for the xenotransplantation.

Piglets produced from cloned blastocysts cultured in vitro with GM-CSF.

In general, pig embryos established by somatic cell nuclear transfer (SCNT) are transferred at the one-cell stage because of suboptimal embryo culture conditions. Improvements in embryo culture can increase the practical application of late embryo transfer. The goal of this study was to evaluate embryos cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro, and to track the in vivo developmental competency of SCNT-derived blastocysts from these GM-CSF embryos. The receptor for GM-CSF was up-regulated in in vitro-produced embryos when compared to in vivo-produced cohorts, but the level decreased when GM-CSF was present. In vitro fertilized (IVF) embryos, supplemented with GM-CSF (2 or 10 ng/ml), showed a higher frequency of development to the blastocyst stage compared to controls. The total cell numbers of the blastocysts also increased with supplementation of GM-CSF. Molecular analysis demonstrates that IVF-derived blastocysts cultured with GM-CSF exhibit less apoptotic activity. Similarly, an increase in development to the blastocyst stage and an increase in the average total-cell number in the blastocysts were observed when SCNT-derived embryos were cultured with either concentration of GM-CSF (2 or 10 ng/ml). When SCNT-derived embryos, cultured with 10 ng/ml GM-CSF, were transferred into six surrogates at Day 6, five of the surrogates became pregnant and delivered healthy piglets. Our findings suggest that supplementation of GM-CSF can provide better culture conditions for IVF- and SCNT-derived embryos, and pig SCNT-derived embryos cultured with GM-CSF in vitro can successfully produce piglets when transferred into surrogates at the blastocyst stage. Thus, it may be practical to begin performing SCNT-derived embryo transfer at the blastocyst stage.

Hydrogen peroxide-induced VCAM-1 expression in pancreatic islets and beta-cells through extracellular Ca2+ influx.

BACKGROUND: The use of porcine islets as alternatives to transplantable human islets is hampered by xenotransplant rejection. To identify molecular mechanisms that would allow subversion of xenoislet rejection, we investigated the role of H2O2 in vascular cell adhesion molecule-1 (VCAM-1) expression by porcine and mouse islets and beta-cell lines. METHODS: Porcine islets were treated with H2O2, tumor necrosis factor alpha, interferon-gamma, interleukin-1beta, and lipopolysaccharide, to assess the effects of inflammatory stimulators on VCAM-1 expression using flow cytometry. The role of Ca2+ in H2O2-induced VCAM-1 expression was investigated in beta-cell lines using an extracellular Ca2+ chelator and Ca2+-depleted media. Furthermore, H2O2-induced VCAM-1 expression was measured in beta-cells, pretreated with inhibitors of protein kinase C, phospholipase D, and phosphatidylinositol-3 kinase/Akt. Finally, H2O2-induced VCAM-1 expression was evaluated in porcine islets and rodent beta-cell lines infected with an adenovirus encoding catalase, a H2O2-removing enzyme. RESULTS: H2O2 was most potent inflammatory stimulator of VCAM-1 expression in porcine islets and had the greatest effect on VCAM-1 expression by beta-cells. Signaling pathway analysis demonstrated that extracellular Ca2+ influx was critical to H2O2-mediated VCAM-1 expression; however, protein kinase C, phospholipase D, and phosphatidylinositol-3 kinase/Akt activation were not required for VCAM-1 expression. Finally, catalase overexpression inhibited H2O2-induced VCAM-1 expression by islets and beta-cell lines. CONCLUSION: An extracellular calcium-dependent H2O2 pathway is the critical mediator of VCAM-1 expression by pancreatic islets and beta-cells. Inhibition of this pathway by catalase overexpression in donor islets can be exploited to protect against xenoislet immune responses.

Coordinated change of a ratio of methylated H3-lysine 4 or acetylated H3 to acetylated H4 and DNA methylation is associated with tissue-specific gene expression in cloned pig.

Various cell types in higher multicellular organisms are genetically homogenous, but are functionally and morphologically heterogeneous due to the differential expression of genes during development, which appears to be controlled by epigenetic mechanisms. However, the exact molecular mechanisms that govern the tissue-specific gene expression are poorly understood. Here, we show that dynamic changes in histone modifications and DNA methylation in the upstream coding region of a gene containing the transcription initiation site determine the tissue-specific gene expression pattern. The tissue-specific expression of the transgene correlated with DNA demethylation at specific CpG sites as well as significant changes in histone modifications from a low ratio of methylated H3- lysine 4 or acetylated H3-lysine 9, 14 to acetylated H4 to higher ratios. Based on the programmed status of transgene silenced in cloned mammalian ear-derived fibroblasts, the transgene could be reprogrammed by change of histone modification and DNA methylation by inhibiting both histone deacetylase and DNA methylation, resulting in high expression of the transgene. These findings indicate that dynamic change of histone modification and DNA methylation is potentially important in the establishment and maintenance of tissue-specific gene expression.